Focal adhesion kinase-dependent regulation of adhesive forces involves vinculin recruitment to focal adhesions.
Focal adhesion kinase-dependent regulation of adhesive forces involves vinculin recruitment to focal adhesions.
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DOI：
10.1042/bc20090104
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发表时间：
2010-01-14
影响因子：
2.7
通讯作者：
García AJ
中科院分区：
文献类型：
--
作者：
Dumbauld DW;Michael KE;Hanks SK;García AJ
粘着斑激酶（Focal adhesion kinase，FAK）是一种重要的非受体酪氨酸激酶，在细胞迁移反应、粘附信号转导和机械力信号转导中起着重要作用。FAK依赖的细胞迁移调控涉及粘着斑的动态转换，以及肌动蛋白细胞骨架聚合和板状伪足突起。虽然FAK在迁移和机械感应反应中的作用已经确定，但FAK对粘附力产生的贡献还不清楚。使用FAK空细胞表达野生型和突变型FAK下的诱导型四环素启动子，我们分析了FAK的作用，在稳态粘附力的产生，使用微图案化的基板和流体动力学粘附试验。FAK表达降低了30%的稳态强度相比，FAK-空细胞。粘着斑激酶表达减少黏着斑蛋白本地化局灶性粘连35%，独立于整合素结合和本地化的talin和桩蛋白的变化。黏着斑蛋白的RNAi敲低消除了粘着力的FAK依赖性差异。黏着斑蛋白定位和粘附力的FAK依赖性变化在人原代成纤维细胞中被证实，FAK被RNAi敲低。自磷酸化Y397和激酶结构域Y 576/Y 577位点是FAK介导的粘附反应所需的差异。我们证明FAK通过调节黏着斑蛋白募集到局灶性粘连来降低稳态粘连强度。这些发现为FAK在细胞与细胞外基质之间的机械相互作用中的作用提供了见解。
Focal adhesion kinase (FAK), an essential non-receptor tyrosine kinase, plays pivotal roles in migratory responses, adhesive signaling, and mechanotransduction. FAK-dependent regulation of cell migration involves focal adhesion turnover dynamics as well as actin cytoskeleton polymerization and lamellipodia protrusion. Whereas roles for FAK in migratory and mechanosensing responses have been established, the contributions of FAK to the generation of adhesive forces are not well understood. Using FAK-null cells expressing wild-type and mutant FAK under an inducible tetracycline promoter, we analyzed the role of FAK in the generation of steady-state adhesive forces using micropatterned substrates and a hydrodynamic adhesion assay. FAK expression reduced steady-state strength by 30% compared to FAK-null cells. FAK expression reduced vinculin localization to focal adhesions by 35% independently from changes in integrin binding and localization of talin and paxillin. RNAi knockdown of vinculin abrogated the FAK-dependent differences in adhesive force. FAK-dependent changes in vinculin localization and adhesive force were confirmed in human primary fibroblasts with FAK knocked down by RNAi. The autophosphorylation Y397 and kinase domain Y576/Y577 sites were differentially required for FAK-mediated adhesive responses. We demonstrate that FAK reduces steady-state adhesion strength by modulating vinculin recruitment to focal adhesions. These findings provide insights into the role of FAK in mechanical interactions between a cell and the extracellular matrix.